bioRxivPlant Biology
Improving the accuracy of single turnover active fluorometry (STAF) for the estimation of phytoplankton primary productivity (PhytoPP)
Abstract
Photosystem II (PSII) photochemistry is the ultimate source of reducing power for phytoplankton primary productivity (PhytoPP). Single turnover active chlorophyll fluorometry (STAF) provides a non-intrusive method that has the potential to measure PhytoPP on much wider spatiotemporal scales than is possible with more direct methods such as 14C fixation and O2 evolved through water oxidation. Application of a STAF-derived absorption coefficient for PSII light-harvesting (aLHII) provides a method for estimating PSII photochemical flux on a unit volume basis (JVPII). Within this study, we assess potential errors in the calculation of JVPII arising from sources other than photochemically active PSII complexes (baseline fluorescence) and the package effect. Although our data show that such errors can be significant, we identify fluorescence-based correction procedures that can be used to minimize their impact. For baseline fluorescence, the correction incorporates an assumed consensus PSII photochemical efficiency for dark-adapted material. The error generated by the package effect can be minimized through the ratio of variable fluorescence measured within narrow wavebands centered at 730 nm, where the re-absorption of PSII fluorescence emission is minimal, and at 680 nm, where re-absorption of PSII fluorescence emission is maximal. We conclude that, with incorporation of these corrective steps, STAF can provide a reliable estimate of JVPII and, if used in conjunction with simultaneous satellite measurements of ocean color, could take us significantly closer to achieving the objective of obtaining reliable autonomous estimates of PhytoPP.
- assay: fluorescence / live-cell microscopy
- research focus: cell signaling & trafficking
- approach: experimental assay
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